Despite the fact that tuberculosis is a major public global health problem, there have been no drugs developed for it since the 1960s. In 2006 tuberculosis caused 1.7 million deaths annually with approximately 8 million new cases emerging annually. Tuberculosis has been exacerbated by human immunodeficiency virus pandemic with over 50% of deaths among the HIV-infected resulting from co- infection with Mycobacterium tuberculosis. The current treatment regimen of tuberculosis has been unchanged since the 1980s and relies on two months of intensive, directly observed therapy with isoniazid, rifampicin, pyrazinamide, and ethambutol followed by a minimum of four months of isoniazid and rifampicin. One of the key reasons for the long length of a drug regimen is the subpopulation of isolates of M. tuberculosis that exhibit non-replicating persistence (NRP). Thus, screening for compounds against nonreplicating M. tuberculosis may lead to drugs that require a shorter drug regimen. Our goal is to identify novel drugs for the treatment of Tuberculosis. This grant proposal brings together the expertise the Institute for Tuberculosis Research at University of Illinois at Chicago (ITR- UIC), and LifePharms, a natural product drug discovery company. LifePharms possesses a unique and previously unprobed natural product library of prefractionated extracts of North American basidiomycetes and ascomycetes (mushrooms) and expertise in the isolation and structural identification of compounds from this source. We have now screened 29,000 fractions (out of a total of 200,000) and have identified 4 partially purified compounds with both potent activity against M. tuberculosis without being toxic to mammalian cells (selectivity >10). More importantly, two of these compounds are also active against the nonreplicating persistence form of M. tuberculosis. Additionally, we have identified several other partially purified compounds with anti-tuberculosis activity. Here, we propose to a) evaluate all active fractions for activity vs mammalian cells and NPR-TB;b) determine activity against drug resistant TB and SNP cluster representatives with the compounds that meet our activity criteria, C) isolate and chemically identify the compounds with the best activity profile, and D) .Initiate the acquisition of larger amounts of the source organism or develop a synthesis plan for the most attractive compounds. PUBLIC HEALTH RELEVANCE: Despite the fact that tuberculosis is a major public global health problem, there have been no drugs developed for it since the 1960s. Moreover, tuberculosis has been exacerbated by human immunodeficiency virus pandemic with over 50% of deaths among the HIV-infected resulting from co-infection with Mycobacterium tuberculosis. We have discovered anti-TB lead compounds from an unexplored natural product. It is our goal to determine if these compounds represent new therapies for M. tuberculosis and develop them if warranted.